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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

Online
ISSN
1437-4331
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Volume 47, Issue 4

Issues

The selection and application of ssDNA aptamers against MPT64 protein in Mycobacterium tuberculosis

Lianhua Qin
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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/ Ruijuan Zheng
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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/ Zhanzhong Ma
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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/ Yonghong Feng
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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/ Zhonghua Liu
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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/ Hua Yang
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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/ Jie Wang
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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/ Ruiliang Jin
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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/ Junmei Lu
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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/ Yuansheng Ding
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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/ Zhongyi Hu
  • Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated to Tongji University, Shanghai, China
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Published Online: 2009-03-02 | DOI: https://doi.org/10.1515/CCLM.2009.097

Abstract

Background: Tuberculosis (TB) remains a major health problem affecting millions of people worldwide. One-third of the world's population is infected with Mycobacterium tuberculosis, the etiologic agent of TB. A simple and rapid method to diagnose TB is urgently needed to be developed. The procedure of systematic evolution of ligands by exponential enrichment (SELEX) is a method in which single-stranded oligonucleotides (called aptamers) are selected from a wide variety of sequences, based on their interaction with a target molecule. Aptamers have been used in numerous investigations as therapeutic or diagnostic tools.

Methods: In this study, we apply a SELEX method to develop aptamers against MPT64 protein from M. tuberculosis. On this basis, a sandwich assay scheme with the complex of aptamer-MPT64 was designed and tested the feasibility of detecting M. tuberculosis by detecting MPT64 protein levels in the culture filtrates of 77 samples including M. tuberculosis and other Mycobacterium species.

Results: There was a highly significant difference (p<0.01) between group A (non-TB Mycobacterium, bacille Calmette-Guérin) and group B (M. tuberculosis, M. bovis), when they were diagnosed with the sandwich assay scheme based on aptamer-protein complex to detect MPT64 protein levels in the culture filtrates of samples. When the cut-off point was at the optical density value of 0.58 (95%=0.764–0.946; Z=6.130, p=0.0001), the sandwich assay scheme based on aptamer-protein complex had a high sensitivity (negative ration, 24/27, 86.3%) and specificity (positive ration, 46/52, 88.5%).

Conclusions: Aptamer of MPT64 as a new detection tool, to a certain extent, is feasible to diagnose Mycobacterium tuberculosis.

Clin Chem Lab Med 2009;47:405–11.

Keywords: aptamer; aptamer-protein sandwich assay scheme; MPT64 protein; Mycobacterium tuberculosis; systematic evolution of ligands by exponential enrichment (SELEX)

About the article

Corresponding author: Zhongyi Hu, Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital Affiliated to Tongji University, 507 Zhengmin Road, Shanghai, 200433, China Phone: +86-21-65115006 ext. 3037, Fax: +86-21-65111298,


Received: 2008-11-10

Accepted: 2009-01-06

Published Online: 2009-03-02

Published in Print: 2009-04-01


Citation Information: Clinical Chemistry and Laboratory Medicine, Volume 47, Issue 4, Pages 405–411, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2009.097.

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